Detection of changes in avian influenza genome fragments by multitemperature single-strand conformational polymorphism.
We have applied the temperature-dependent single-stranded conformational polymorphism (SSCP) analysis for characterization of influenza A virus cDNA fragments. A series of primers were synthesized on the basis of the comparison of hemagglutinin and PB2 gene sequences of different origin. RT-PCR reactions were run using these primers, products were denatured and single-strand DNA fragments were subjected to electrophoresis. After silver staining, ssDNA band patterns characteristic for each genotype were observed. Minor differences within a serotype were detected without need for sequencing the PCR products making the preliminary analysis of avian influenza isolates much faster.
Gromadzka
Beata
B
Department of Molecular Virology, Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Kladki 24, Gdansk, Poland.
Smietanka
Krzysztof
K
Dragun
Jakub
J
Minta
Zenon
Z
Gora-Sochacka
Anna
A
Szewczyk
Boguslaw
B
D000818:Animals; D001717:Birds / Q000821:virology; D018076:DNA, Complementary / Q000235:genetics; D016679:Genome, Viral; D005838:Genotype; D006801:Humans; D009980:Influenza A virus / Q000235:genetics*; D005585:Influenza in Birds / Q000145:classification / Q000175:diagnosis / Q000235:genetics; D010802:Phylogeny; D018807:Polymorphism, Single-Stranded Conformational; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013696:Temperature
D018076:DNA, Complementary